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. 2011 Sep 1;22(17):3055–3069. doi: 10.1091/mbc.E11-01-0069

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© 2011 Xiao et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 7: — Colocalization of CDK4 mRNA with PBs after miR-222 overexpression and following polyamine depletion. (A) Schematic of the plasmids used for the visualization of CDK4 mRNA. PMS2 and pMS2-CDK4 expressed MS2 and MS2-CDK4 RNAs, each containing 12 tandem MS2 hairpins; pMS2-YFP expressed a fusion fluorescent protein (MS2-YFP) capable of detecting MS2-containing RNA. (B) Images of CDK4 mRNA colocalization with PBs in cells overexpressing miR-222. Using confocal microscopy, MS2 and MS2-CDK4 mRNA were visualized using MS2-YFP (green fluorescence); red, RCK (PB marker) signals; yellow, colocalized red and green signals. (C) Images of CDK4 mRNA colocalization with PBs in polyamine-deficient cells after CUGBP1 silencing or silencing both CUGBP1 and miR-222. CDK4 mRNA was detected using pMS2-CDK4 as explained in (B); three experiments were performed that showed similar results.